miR494对结直肠癌细胞迁移能力的影响及其分子机制

Effect of miR494 on migration of colorectal cancer cells and its molecular mechanism

目的探究miR494对结直肠癌细胞迁移能力的影响及其分子机制。方法采用慢病毒包装系统建立miR494-Lovo细胞株(转染miR494前体载体)和NC-Lovo细胞株(转染对照载体)。采用定量RT-PCR检测转染率和上皮间质转化标志物E-cadherin、N-cadherin、matrix metallopeptidase 2(MMP2)、MMP9、vimentin和snail表达情况。通过transwell和划痕实验检测细胞迁移能力。结果与NC-Lovo细胞比较,miR494-Lovo细胞中miR494高表达(P<0.05)。与NC-Lovo细胞比较,转染24 h后,miR494-Lovo细胞的迁移能力增强(P<0.05)。转染48 h后,miR494-Lovo细胞E-cadherin表达升高,N-cadherin、MMP2、MMP9、vimentin和snail表达下降(均P<0.05)。结论miR494促进结直肠癌细胞迁移能力,并可能与EMT发生有关。

Objective To investigate the molecular mechanism of miR494 in the migration of colorectal cancer cells.Methods Lentivirus packaging system was used to construct miR494-Lovo cell line with pre-miR494 vector transfection and NC-Lovo cell line with control vector transfection,and fluorescence quantitative RT-PCR was used to detect the transfection rate.Cell migration was detected by transwell and scratch test,and the levels of epithelial-mesenchymal transition(EMT)related proteins,including E-cadherin,N-cadherin,matrix metallopeptidase 2(MMP2),MMP9,vimentin and snail,were detected by RT-PCR.Results miR494 was highly expressed in miR494-Lovo cells as compared with NC-Lovo cells(P<0.05).After 24 h transfection,the migration ability of miR494-Lovo cells was enhanced compared with that of NC-Lovo cells(P<0.05).After 48 h transfection,the expression of E-cadherin increased and the levels of N-cadherin,MMP2,MMP9,vimentin and snail decreased in miR494-Lovo cells(all P<0.05).Conclusions miR494 can promote the occurrence of EMT in colorectal cancer and increase the migration ability of colorectal cancer cells.